Phenylnaphthalene hydroxy acids

ABSTRACT

Compounds having the formula I:    &lt;IMAGE&gt;  I  are inhibitors of leukotriene biosynthesis. These compounds are useful as anti-asthmatic, anti-allergic, anti-inflammatory, and cytoprotective agents. They are also useful in treating angina, cerebral spasm, glomerular nephritis, hepatitis, endotoxemia, uveitis, and allograft rejection and in preventing the formation of atherosclerotic plaques.

BACKGROUND OF THE INVENTION

The leukotrienes constitute a group of locally acting hormones, producedin living systems from arachidonic acid. The major leukotrienes areLeukotriene B₄ (abbreviated at LTB₄), LTC₄, LTD₄, and LTE₄. Thebiosynthesis of these leukotrienes begins with the action of the enzyme5-lipoxygenase on arachidonic acid to produce the epoxide known asLeukotriene A₄ (LTA₄), which is converted to the other leukotrienes bysubsequent enzymatic steps. Further details of the biosynthesis as wellas the metabolism of the leukotrienes are to be found in the bookLeukotrienes and Lipoxygenases, ed. J. Rokach, Elsevier, Amsterdam(1989). The actions of the leukotrienes in living systems and theircontribution to various diseases states are also discussed in the bookby Rokach.

European patent application 375,404 (Jun. 27, 1990) describes certainnaphthalene-containing heterocyclic ethers of structure A which areinhibitors of the enzyme 5-lipoxygenase. EP application 375,452 (Jun.27, 1990) describes naphthalene-containing hydrocarbon ethers ofstructure B which are reported to possess the same activity. EPapplication 462,830 (Dec. 27, 1991) describes bicyclicheterocycle-containing hydrocarbon ethers of structure C which arereported to possess the same activity. All these series of prior artcompounds differ significantly from the present invention in that theylack the aryl substituent of the present compounds.

A series of natural products known as the justicidins are referred to inthe Merck Index, 11^(th) edition, 1989, no. 5154. The justicidins differconsiderably from the present compounds in that they are lacking thelarge Het group.

Hence the compounds of the present invention are completely novel andunexpectedly have biological activity as leukotriene biosynthesisinhibitors. ##STR2##

SUMMARY OF THE INVENTION

The present invention relates to phenylnaphthalene hydroxy acids havingactivity as leukotriene biosynthesis inhibitors, to methods for theirpreparation, and to methods and pharmaceutical formulations for usingthese compounds in mammals (especially humans).

Because of their activity as leukotriene biosynthesis inhibitors, thecompounds of the present invention are useful as anti-asthmatic,anti-allergic, anti-inflammatory, and cytoprotective agents. They arealso useful in treating angina, cerebral spasm, glomerular nephritis,hepatitis, endotoxemia, uveitis, and allograft rejection and inpreventing the formation of atherosclerotic plaques.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the present invention may be represented by thefollowing formula I: ##STR3## wherein: R¹, R⁵, and R¹¹ is eachindependently H, OH, lower alkyl, or lower alkoxy;

R² is H, lower alkyl, or together with R¹ forms a double bonded oxygen(═O);

R³ is H, lower alkyl, hydroxy lower alkyl, lower alkoxy lower alkyl, oris joined to R¹ to form a carbon bridge of 2 or 3 carbon atoms or amono-oxa carbon bridge of 1 or 2 carbon atoms, said bridge optionallycontaining a double bond;

R⁴, R¹², and R¹⁴ is each independently H or lower alkyl;

R⁶ is H or lower alkyl, or two R⁶ groups on the same or adjacent carbonscan form a saturated ring of 3 to 8 members;

R⁷ is H, OH, lower alkyl, lower alkoxy, lower alkylthio, or loweralkylcarbonyloxy;

R⁸ is H, halogen, lower alkyl, lower alkoxy, CF₃, CN, COR¹⁴, or anon-bonded electron pair;

R⁹ and R¹⁰ is each independently H, lower alkyl, lower alkoxy, hydroxylower alkyl, lower alkoxy lower alkyl, lower alkylthio lower alkyl,lower alkythio lower alkylcarbonyl, (R⁸)₂ -phenylthio lower alkyl,halogen, CN, NO₂, CF₃, N₃, N(R¹³)₂, NR¹³ COR¹⁴, NR¹³ CON(R¹³)₂, SR¹⁵,S(O)R¹⁵, S(O)₂ R¹⁵, S(O)₂ N(R¹³)₂, COR¹⁴, CON(R¹³)₂, CO₂ R¹⁴, C(R¹⁴)₂OC(R¹⁴)₂ --CO₂ R¹⁴, or C(R¹⁴)₂ CN;

R¹⁰ is attached to either ring of the naphthalene ring system;

R¹³ is H or lower alkyl, or two R¹³ groups attached to the same nitrogenmay form a saturated ring of 5 or 6 members, optionally containing asecond heteroatom chosen from O, S, or NR⁴ ;

R¹⁵ is lower alkyl, phenyl-(R⁸)₂, or CF₃ ;

X¹ is O, S, S(O), S(O)₂, or C(R⁶)₂ ;

X² is O, S, C(R⁶)₂, or bond;

X³ is C(R⁶)₂ S, SC(R⁶)₂, C(R⁶)₂ O, OC(R⁶)₂, CR⁶ ═CR⁶, C(R⁶)₂ C(R⁶)₂, O,or S;

Ar is phenyl(R⁹)₂ or naphthyl(R⁹)₂ ;

Het is arylene-(R⁸)₂, wherein arylene is a 5-membered aromatic ringwherein one carbon atom is replaced by O or S and 0-2 carbon atoms arereplaced by N; a 5-membered aromatic ring where 1-3 carbon atoms arereplaced by N; a 6-membered aromatic ring wherein 1-3 carbon atoms arereplaced by N; 2- or 4-pyranone; or 2- or 4-pyridinone;

m is 0 or 1

or a pharmaceutically acceptable salt thereof.

It will be obvious to one skilled in the art that if R¹¹ is OH or loweralkoxy on the same carbon atom as the OH group of I, the elements ofwater or an alcohol may be lost to give an aldehyde or ketone.

A preferred embodiment of the present invention is represented byFormula Ia: ##STR4## wherein the substituents are as defined for FormulaI, or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is represented by Formula Ib##STR5## wherein: R¹ is H, Me, OMe;

R³ is H or Me;

R¹ and R³ together are --CH═CH--, --CH₂ CH₂ --, --OCH₂ --, or --CH₂)--;

R⁷ is OH or OMe; and

Het is 5,3-Pye; 6,2-Pye; 4,2-Pye, or 2,4-Pye;

or a pharmaceutically acceptable salt thereof.

Definitions

The following abbreviations have the indicated meanings:

Ac=acetyl

Bn=benzyl

c-Bu=cyclobutyl

c-Pen=cyclopentyl

c-Pr=cyclopropyl

c-Hex=cyclohexyl

i-Pt=isopropyl

n-Pr=normal propyl

n-Bu=normal butyl

i-Bu=isobutyl

s-Bu=secondary butyl

t-Bu=tertiary butyl

Et=ethyl

Me=methyl

Ph=phenyl

Pye=pyridindiyl

Thi=thiophenediyl

C₃ H₅ =allyl

Alkyl is intended to include linear and branched structures andcombinations thereof.

The term "alkyl" includes "lower alkyl" and extends to cover carbonfragments having up to 20 carbon atoms. Examples of alkyl groups includeoctyl, nonyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl,eicosyl, 3,7-diethyl-2,2-dimethyl-4-propylnonyl, and the like.

"Lower alkyl" means alkyl groups of from 1 to 7 carbon atoms. Examplesof lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl,s- and t-butyl, pentyl, hexyl, heptyl, and the like.

"Lower alkoxy" means alkoxy groups of from 1 to 7 carbon atoms of astraight, branched, or cyclic configuration. Examples of lower alkoxygroups include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy,cyclohexyloxy, and the like.

"Hydroxy lower alkyl" means a lower alkyl group carrying a hydroxygroup; e.g. --CH₂ CH(OH)CH₂ CH₃.

"Lower alkoxy lower alkyl" means a lower alkyl group carrying a loweralkoxy group; e.g. --CH₂ CH₂ OCH₃.

"Lower alkylthio" means alkylthio groups of from 1 to 7 carbon atoms ofa straight, branched, or cyclic configuration. Examples of loweralkylthio groups include methylthio, ethylthio, isopropylthio,cyclobutylthio, and the like.

"Lower alkylcarbonyl" means alkylcarbonyl groups of from 1 to 8 carbonatoms of a straight, branched, or cyclic configuration. Examples oflower alkylcarbonyl groups are formyl, 2-methylbutanoyl,cyclohexylacetyl, etc. By way of illustration, the 2-methylbutanoylgroup signifies --COCH(CH₃)CH₂ CH₃.

Examples of "arylene" are furan, thiophene, isoxazole, isothiazole,oxazole, thiazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole,1,2,5-oxadiazole, 1,2,5-thiadiazole, pyrrole, pyrazole, imidazole,1,3,4-triazole, pyridine, pyrazine, pyrimidine, pyridazine,1,2,3-triazine, 1,2,4-triazine, 1,3,5-triazine, and the like.

Halogen includes F, Cl, Br, and I.

It is intended that the definitions of any substituent (e.g., R⁴, R⁸,etc.) in a particular molecule be independent of its definitionselsewhere in the molecule. Thus, --C(R⁶)₂ O-- represents --CHCH₃ O--,--C(CH₃)₂ O--, etc.

Optical Isomers--Diastereomers--Geometric Isomers

Some of the compounds described herein contain one or more asymmetriccenters and may thus give rise to diastereomers and optical isomers. Thepresent invention is meant to comprehend such possible diastereomers aswell as their racemic and resolved, enantiomerically pure forms andpharmaceutically acceptable salts thereof.

Some of the compounds described herein contain olefinic double bonds,and unless specified otherwise, are meant to include both E and Zgeometric isomers.

Salts

The pharmaceutical compositions of the present invention comprise acompound of Formula I as an active ingredient or a pharmaceuticallyacceptable salt, thereof, and may also contain a pharmaceuticallyacceptable carrier and optionally other therapeutic ingredients. Theterm "pharmaceutically acceptable salts" refers to salts prepared frompharmaceutically acceptable non-toxic bases including inorganic basesand organic bases. Salts derived from inorganic bases include aluminum,ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganicsalts, manganous, potassium, sodium, zinc and the like. Particularlypreferred are the ammonium, calcium, magnesium, potassium and sodiumsalts. Salts derived from pharmaceutically acceptable organic non-toxicbases include salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as arginine, betaine,caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine, and the like.

When the compound of the present invention is basic, salts may beprepared from pharmaceutically acceptable non-toxic acids, includinginorganic and organic acids. Such acids include acetic, benzenesulfonic,benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic,glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and thelike. Particularly preferred are citric, hydrobromic, hydrochloric,maleic, phosphoric, sulfuric, and tartaric acids.

It will be understood that in the discussion of methods of treatmentwhich follows, references to the compounds of Formula I are meant toalso include the pharmaceutically acceptable salts.

Utilities

The ability of the compounds of Formula I to inhibit biosynthesis of theleukotrienes makes them useful for preventing or reversing the symptomsinduced by the leukotrienes in a human subject. This inhibition of themammalian biosynthesis of leukotrienes indicates that the compounds andpharmaceutical compositions thereof are useful to treat, prevent, orameliorate in mammals and especially in humans: 1) pulmonary disordersincluding diseases such as asthma, chronic bronchitis, and relatedobstructive airway diseases, 2) allergies and allergic reactions such asallergic rhinitis, contact dermatitis, allergic conjunctivitis, and thelike, 3) inflammation such as arthritis or inflammatory bowel disease,4) pain, 5) skin disorders such as psoriasis, atopic eczema, and thelike, 6) cardiovascular disorders such as angina, formation ofatherosclerotic plaques, myocardial ischemia, hypertension, plateletaggregation, and the like, 7) renal insufficiency arising from ischaemiainduced by immunological or chemical (cyclosporin) etiology, 8) migraineor cluster headache, 9) ocular conditions such as uveitis, 10) hepatitisresulting from chemical, immunological, or infectious stimuli, 11)trauma or shock states such as burn injuries, endotoxemia, and the like,12) allograft rejection, 13) prevention of side effects associated withtherapeutic administration of cytokines such as Interleukin II and tumornecrosis factor, 14) chronic lung diseases such as cystic fibrosis,bronchitis, and other small- and large-airway diseases, 15)cholecystitis, and 16 ) multiple sclerosis.

Thus, the compounds of the present invention may also be used to treator prevent mammalian (especially, human) disease states such as erosivegastritis; erosive esophagitis; diarrhea; cerebral spasm; prematurelabor; spontaneous abortion; dysmenorrhea; ischemia; noxiousagent-induced damage or necrosis of hepatic, pancreatic, renal, ormyocardial tissue; liver parenchymal damage caused by hepatoxic agentssuch as CCl₄ and D-galactosamine; ischemic renal failure;disease-induced hepatic damage; bile salt induced pancreatic or gastricdamage; trauma- or stress-induced cell damage; and glycerol-inducedrenal failure. The compounds also act as inhibitors of tumor metastasisand exhibit cytoprotective action.

The cytoprotective activity of a compound may be observed in bothanimals and man by noting the increased resistance of thegastrointestinal mucosa to the noxious effects of strong irritants, forexample, the ulcerogenic effects of aspirin or indomethacin. In additionto lessening the effect of non-steroidal anti-inflammatory drugs on thegastrointestinal tract, animal studies show that cytoprotectivecompounds will prevent gastric lesions induced by oral administration ofstrong acids, strong bases, ethanol, hypertonic saline solutions and thelike.

Two assays can be used to measure cytoprotective ability. These assaysare; (A) an ethanol-induced lesion assay and (B) an indomethacin-inducedulcer assay and are described in EP 140,684.

Dose Ranges

The magnitude of prophylactic or therapeutic dose of a compound ofFormula I will, of course, vary with the nature of the severity of thecondition to be treated and with the particular compound of Formula Iand its route of administration. It will also vary according to the age,weight and response of the individual patient. In general, the dailydose range for anti-asthmatic, anti-allergic or anti-inflammatory useand generally, uses other than cytoprotection, lie within the range offrom about 0.001 mg to about 100 mg per kg body weight of a mammal,preferably 0.01 mg to about 10 mg per kg, and most preferably 0.1 to 1mg per kg, in single or divided doses. On the other hand, it may benecessary to use dosages outside these limits in some cases.

For use where a composition for intravenous administration is employed,a suitable dosage range for anti-asthmatic, anti-inflammatory oranti-allergic use is from about 0.001 mg to about 25 mg (preferably from0.01 mg to about 1 mg) of a compound of Formula I per kg of body weightper day and for cytoprotective use from about 0.1 mg to about 100 mg(preferably from about 1 mg to about 100 mg and more preferably fromabout 1 mg to about 10 mg) of a compound of Formula I per kg of bodyweight per day.

In the case where an oral composition is employed, a suitable dosagerange for anti-asthmatic, anti-inflammatory or anti-allergic use is,e.g. from about 0.01 mg to about 100 mg of a compound of Formula I perkg of body weight per day, preferably from about 0.1 mg to about 10 mgper kg and for cytoprotective use from 0.1 mg to about 100 mg(preferably from about 1 mg to about 100 mg and more preferably fromabout 10 mg to about 100 mg) of a compound of Formula I per kg of bodyweight per day.

For the treatment of diseases of the eye, ophthalmic preparations forocular administration comprising 0.001-1% by weight solutions orsuspensions of the compounds of Formula I in an acceptable ophthalmicformulation may be used.

The exact amount of a compound of the Formula I to be used as acytoprotective agent will depend on, inter alia, whether it is beingadministered to heal damaged cells or to avoid future damage, on thenature of the damaged cells (e.g., gastrointestinal ulcerations vs.nephrotic necrosis), and on the nature of the causative agent. Anexample of the use of a compound of the Formula I in avoiding futuredamage would be co-administration of a compound of the Formula I with anon-steroidal anti-inflammatory drug that might otherwise cause suchdamage (for example, indomethacin). For such use, the compound ofFormula I is administered from 30 minutes prior up to 30 minutes afteradministration of the NSAID. Preferably it is administered prior to orsimultaneously with the NSAID, (for example, in a combination dosageform).

Pharmaceutical Compositions

Any suitable route of administration may be employed for providing amammal, especially a human with an effective dosage of a compound of thepresent invention. For example, oral, rectal, topical, parenteral,ocular, pulmonary, nasal, and the like may be employed. Dosage formsinclude tablets, troches, dispersions, suspensions, solutions, capsules,creams, ointments, aerosols, and the like.

The pharmaceutical compositions of the present invention comprise acompound of Formula I as an active ingredient or a pharmaceuticallyacceptable salt thereof, and may also contain a pharmaceuticallyacceptable carrier and optionally other therapeutic ingredients. Theterm "pharmaceutically acceptable salts" refers to salts prepared frompharmaceutically acceptable non-toxic bases or acids including inorganicbases or acids and organic bases or acids.

The compositions include compositions suitable for oral, rectal,topical, parenteral (including subcutaneous, intramuscular, andintravenous), ocular (ophthalmic), pulmonary (nasal or buccalinhalation), or nasal administration, although the most suitable routein any given case will depend on the nature and severity of theconditions being treated and on the nature of the active ingredient.They may be conveniently presented in unit dosage form and prepared byany of the methods well-known in the art of pharmacy.

For administration by inhalation, the compounds of the present inventionare conveniently delivered in the form of an aerosol spray presentationfrom pressurized packs or nebulisers. The compounds may also bedelivered as powders which may be formulated and the powder compositionmay be inhaled with the aid of an insufflation powder inhaler device.The preferred delivery system for inhalation is a metered doseinhalation (MDI) aerosol, which may be formulated as a suspension orsolution of compound I in suitable propellants, such as fluorocarbons orhydrocarbons.

Suitable topical formulations of Compound I include transdermal devices,aerosols, creams, ointments, lotions, dusting powders, and the like.

In practical use, the compounds of Formula I can be combined as theactive ingredient in intimate admixture with a pharmaceutical carrieraccording to conventional pharmaceutical compounding techniques. Thecarrier may take a wide variety of forms depending on the form ofpreparation desired for administration, e.g., oral or parenteral(including intravenous). In preparing the compositions for oral dosageform, any of the usual pharmaceutical media may be employed, such as,for example, water, glycols, oils, alcohols, flavoring agents,preservatives, coloring agents and the like in the case of oral liquidpreparations, such as, for example, suspensions, elixirs and solutions;or carriers such as starches, sugars, microcrystalline cellulose,diluents, granulating agents, lubricants, binders, disintegrating agentsand the like in the case of oral solid preparations such as, forexample, powders, capsules and tablets, with the solid oral preparationsbeing preferred over the liquid preparations. Because of their ease ofadministration, tablets and capsules represent the most advantageousoral dosage unit form in which case solid pharmaceutical carriers areobviously employed. If desired, tablets may be coated by standardaqueous or nonaqueous techniques.

In addition to the common dosage forms set out above, the compounds ofFormula I may also be administered by controlled release means and/ordelivery devices such as those described in U.S. Pat. Nos. 3,845,770;3,916,899; 3,536,809; 3,598,123; 3,630,200, and 4,008,719, thedisclosures of which are hereby incorporated herein by reference.

Pharmaceutical compositions of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient, as a powder or granules or as a solution or a suspension inan aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or awater-in-oil liquid emulsion. Such compositions may be prepared by anyof the methods of pharmacy but all methods include the step of bringinginto association the active ingredient with the carrier whichconstitutes one or more necessary ingredients. In general, thecompositions are prepared by uniformly and intimately admixing theactive ingredient with liquid carriers or finely divided solid carriersor both, and then, if necessary, shaping the product into the desiredpresentation. For example, a tablet may be prepared by compression ormolding, optionally with one or more accessory ingredients. Compressedtablets may be prepared by compressing in a suitable machine, the activeingredient in a free-flowing form such as powder or granules, optionallymixed with a binder, lubricant, inert diluent, surface active ordispersing agent. Molded tablets may be made by molding in a suitablemachine, a mixture of the powdered compound moistened with an inertliquid diluent. Desirably, each tablet contains from about 2.5 mg toabout 500 mg of the active ingredient and each cachet or capsulecontains from about 2.5 to about 500 mg of the active ingredient.

The following are examples of representative pharmaceutical dosage formsfor the compounds of Formula I:

    ______________________________________                                        Injectable Suspension (I.M.)                                                                           mg/mL                                                ______________________________________                                        Compound of Formula I    10                                                   Methylcellulose          5.0                                                  Tween 80                 0.5                                                  Benzyl alcohol           9.0                                                  Benzalkonium chloridel.0                                                      Water for injection to a total volume of 1 mL                                 ______________________________________                                        Tablet                   mg/tablet                                            ______________________________________                                        Compound of Formula I    25                                                   Microcrystalline Cellulose                                                                             415                                                  Povidone                 14.0                                                 Pregelatinized Starch    43.5                                                 Magnesium Stearate       2.5                                                                           500                                                  ______________________________________                                        Capsule                  mg/capsule                                           ______________________________________                                        Compound of Formula I    25                                                   Lactose Powder           573.5                                                Magnesium Stearate       1.5                                                                           600                                                  ______________________________________                                        Aerosol                  Per canister                                         ______________________________________                                        Compound of Formula I      24 mg                                              Lecithin, NF Liquid Concentrate                                                                         1.2 mg                                              Trichlorofluoromethane, NF                                                                             4.025 gm                                             Dichlorodifluoromethane, NF                                                                            12.15 gm                                             ______________________________________                                    

Combinations with other drugs

In addition to the compounds of Formula I, the pharmaceuticalcompositions of the present invention can also contain other activeingredients, such as cyclooxygenase inhibitors, non-steroidalanti-inflammatory drugs (NSAIDs), peripheral analgesic agents such aszomepirac diflunisal and the like. The weight ratio of the compound ofthe Formula I to the second active ingredient may be varied and willdepend upon the effective dose of each ingredient. Generally, aneffective dose of each will be used. Thus, for example, when a compoundof the Formula I is combined with an NSAID the weight ratio of thecompound of the Formula I to the NSAID will generally range from about1000:1 to about 1:1000, preferably about 200:1 to about 1:200.Combinations of a compound of the Formula I and other active ingredientswill generally also be within the aforementioned range, but in eachcase, an effective dose of each active ingredient should be used.

NSAIDs can be characterized into five groups:

(1) the propionic acid derivatives;

(2) the acetic acid derivatives;

(3) the fenamic acid derivatives;

(4) the oxicams; and

(5) the biphenylcarboxylic acid derivatives;

or a pharmaceutically acceptable salt thereof.

The propionic acid derivatives which may be used comprise: alminoprofen,benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen,flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen,oxaprozin, pirprofen, prano-profen, suprofen, tiaprofenic acid, andtioxaprofen. Structurally related propionic acid derivatives havingsimilar analgesic and anti-inflammatory properties are also intended tobe included in this group.

Thus, "propionic acid derivatives" as defined herein are non-narcoticanalgesics/non-steroidal anti-inflammatory drugs having a free--CH(CH₃)COOH or --CH₂ CH₂ COOH group (which optionally can be in theform of a pharmaceutically acceptable salt group, e.g., --CH(CH₃)COO⁻Na⁺ or --CH₂ CH₂ COO⁻ Na⁺), typically attached directly or via acarbonyl function to a ring system, preferably to an aromatic ringsystem.

The acetic acid derivatives which may be used comprise: indomethacin,which is a preferred NSAID, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, andzomepirac. Structually related acetic acid derivatives having similaranalgesic and anti-inflammatory properties are also intended to beencompassed by this group.

Thus, "acetic acid derivatives" as defined herein are non-narcoticanalgesics/non-steroidal anti-inflammatory drugs having a free --CH₂COOH group (which optionally can be in the form of a pharmaceuticallyacceptable salt group, e.g. --CH₂ COO⁻ Na⁺), typically attached directlyto a ring system, preferably to an aromatic or heteroaromatic ringsystem.

The fenamic acid derivatives which may be used comprise: flufenamicacid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamicacid. Structurally related fenamic acid derivatives having similaranalgesic and anti-inflammatory properties are also intended to beencompassed by this group.

Thus, "fenamic acid derivatives" as defined herein are non-narcoticanalgesics/non-steroidal anti-inflammatory drugs which contain the basicstructure: ##STR6## which can bear a variety of substituents and inwhich the free --COOH group can be in the form of a pharmaceuticallyacceptable salt group, e.g., --COO⁻ Na⁺.

The biphenylcarboxylic acid derivatives which can be used comprise:diflunisal and flufenisal. Structurally related biphenylcarboxylic acidderivatives having similar analgesic and anti-inflammatory propertiesare also intended to be encompassed by this group.

Thus, "biphenylcarboxylic acid derivatives" as defined herein arenon-narcotic analgesics/non-steroidal anti-inflammatory drugs whichcontain the basic structure: ##STR7## which can bear a variety ofsubstituents and in which the free --COOH group can be in the form of apharmaceutically acceptable salt group, e.g., --COO⁻ Na⁺.

The oxicams which can be used in the present invention comprise:isoxicam, piroxicam, sudoxicam and tenoxican. Structurally relatedoxicams having similar analgesic and anti-inflammatory properties arealso intended to be encompassed by this group.

Thus, "oxicams" as defined herein are non-narcoticanalgesics/non-steroidal anti-inflammatory drugs which have the generalformula: ##STR8## wherein R is an aryl or heteroaryl ring system.

The following NSAIDs may also be used: amfenac sodium, aminoprofen,anitrazafen, antrafenine, auranofin, bendazac lysinate, benzydanine,beprozin, broperamole, bufezolac, cinmetacin, ciproquazone, cloximate,dazidamine, deboxamet, delmetacin, detomidine, dexindoprofen, diacerein,di-fisalamine, difenpyramide, emorfazone, enfenamic acid, enolicam,epirizole, etersalate, etodolac, etofenamate, fanetizole mesylate,fenclorac, fendosal, fenflumizole, feprazone, floctafenine, flunixin,flunoxaprofen, fluproquazone, fopirtoline, fosfosal, furcloprofen,glucametacin, guaimesal, ibuproxam, isofezolac, isonixim, isoprofen,isoxicam, lefetamine HCl, leflunomide, lofemizole, lonazolac calcium,lotifazole, loxoprofen, lysin clonixinate, meclofenamate sodium,meseclazone, nabumetone, nictindole, nimesulide, orpanoxin, oxametacin,oxapadol, perisoxal citrate, pimeprofen, pimetacin, piproxen, pirazolac,pirfenidone, proglumetacin maleate, proquazone, pyridoxiprofen,sudoxicam, talmetacin, talniflumate, tenoxicam, thiazolinobutazone,thielavin B, tiaramide HCl, tiflamizole, timegadine, tolpadol,tryptamid, and ufenamate.

The following NSAIDs, designated by company code number (see e.g.,Pharmaprojects), may also be used: 480156S, AA861, AD1590, AFP802,AFP860, AI77B, AP504, AU8001, BPPC, BW540C, CHINOIN 127, CN100, EB382,EL508, F1044, GV3658, ITF182, KCNTEI6090, KME4, LA2851, MR714, MR897,MY309, ONO3144, PR823, PV102, PV108, R830, RS2131, SCR152, SH440,SIR133, SPAS510, SQ27239, ST281, SY6001, TA60, TAI-901(4-benzoyl-1-indancarboxylic acid), TVX2706, U60257, UR2301, andWY41770.

Finally, NSAIDs which may also be used include the salicylates,specifically acetyl salicylic acid and the phenylbutazones, andpharmaceutically acceptable salts thereof.

In addition to indomethacin, other preferred NSAIDS are acetyl salicylicacid, diclofenac, fenbufen, fenoprofen, flurbiprofen, ibuprofen,ketoprofen, naproxen, phenylbutazone, piroxicam, sulindac and tolmetin.

Pharmaceutical compositions comprising the Formula I compounds may alsocontain inhibitors of the biosynthesis of the leukotrienes such as aredisclosed in EP 138,481 (Apr. 24, 1985), EP 115,394 (Aug. 8, 1984), EP136,893 (Apr. 10, 1985), and EP 140,709 (May 8, 1985), which are herebyincorporated herein by reference.

The compounds of the Formula I may also be used in combination withleukotriene antagonists such as those disclosed in EP 106,565 (Apr. 25,1984) and EP 104,885 (Apr. 4, 1984) which are hereby incorporated hereinby reference and others known in the art such as those disclosed in EPApplication Nos. 56,172 (Jul. 21, 1982) and 61,800 (Jun. 10, 1982); andin U.K. Patent Specification No. 2,058,785 (Apr. 15, 1981), which arehereby incorporated herein by reference.

Pharmaceutical compositions comprising the Formula I compounds may alsocontain as the second active ingredient, prostaglandin antagonists suchas those disclosed in EP 11,067 (May 28, 1980) or thromboxaneantagonists such as those disclosed in U.S. Pat. No. 4,237,160. They mayalso contain histidine decarboxylase inhibitors such asα-fluoromethylhistidine, described in U.S. Pat. No. 4,325,961. Thecompounds of the Formula I may also be advantageously combined with anH₁ - or H₂ -receptor antagonist, such as for instance acetamazole,aminothiadiazoles disclosed in EP 40,696 (Dec. 2, 1981), benadryl,cimetidine, famotidine, framamine, histadyl, phenergan, ranitidine,terfenadine and like compounds, such as those disclosed in U.S. Pat.Nos. 4,283,408; 4,362,736; and 4,394,508. The pharmaceuticalcompositions may also contain a K⁺ /H⁺ ATPase inhibitor such asomeprazole, disclosed in U.S. Pat. No. 4,255,431, and the like.Compounds of Formula I may also be usefully combined with most cellstabilizing agents, such as1,3-bis(2-carboxychromon-5-yloxy)-2-hydroxypropane and related compoundsdescribed in British Patent Specifications 1,144,905 and 1,144,906.Another useful pharmaceutical composition comprises the Formula Icompounds in combination with serotonin antagonists such asmethysergide, the serotonin antagonists described in Nature, Vol. 316,pages 126-131, 1985, and the like. Each of the references referred to inthis paragraph is hereby incorporated herein by reference.

Other advantageous pharmaceutical compositions comprise the Formula Icompounds in combination with anti-cholinergics such as ipratropiumbromide, bronchodilators such as the beta agonist salbutamol,metaproterenol, terbutaline, fenoterol and the like, and theanti-asthmatic drugs theophylline, choline theophyllinate andenprofylline, the calcium antagonists nifedipine, diltiazem,nitrendipine, verapamil, nimodipine, felodipine, etc. and thecorticosteroids, hydrocortisone, methylprednisolone, betamethasone,dexamethasone, beclomethasone, and the like.

Methods of Synthesis

Compounds of the present invention can be prepared according to thefollowing methods. The substituents are the same as in Formula I exceptwhere defined otherwise.

Scheme I

The route which is used to prepare the lactone VI is outlined in SchemeI. The aryl carboxaldehyde II is converted to the thioacetal III bytreatment with thiophenol in the presence of a Lewis-Acid such asBF₃.Et₂ O in an organic solvent such as isopropyl acetate. Thethioacetal III is then converted to the lactone V by treatment with abase such as n-BuLi in an organic solvent such as THF followed by thesuccessive additions of 2-(5H) furanone and the benzyloxyarylcarboxaldehyde IV in an organic solvent such as THF and after quenchingwith an acid such as HOAc. The cyclization, dehydration anddebenzylation are achieved simultaneously by heating the lactone V inthe presence of an acid such as TFA, and using thioanisole as organicsolvent providing the lactone VI.

Scheme II

The preparation of compounds of Formula I (wherein X³ =--C(R⁶)₂ O--) isdescribed in Scheme II. A first method requires coupling of the naphtholVI with a benzylic halide or activated alcohol of type vII (whereinX=Cl, Br, I, OMs, OTs) in a polar organic solvent such DMF in thepresence of an inorganic base such as Cs₂ CO₃.

In an alternate procedure, the naphthol VI is condensed with theheterocyclic alcohol VII (wherein X=OH) in the presence of a phosphinesuch as Ph₃ P and a azodicarboxylate diester, in a solvent such as THF,to afford Formula I (wherein X³ =--C(R⁶)₂ O--) compounds.

Scheme III

The synthesis of compounds of Formula I (wherein X³ =--OCH₂ --) isdescribed in Scheme III. The phenol lactone VI may be converted to thetriflate VIII by treatment with trifluoromethanesulfonic anhydride inthe presence of an organic base such as pyridine in a solvent such asCH₂ Cl₂. Subsequent treatment of VIII in a solvent such as DMSO/MeOHwith an organic base such as triethylamine, a phosphine such as1,1'-bis(diphenylphosphino)ferrocene, a palladium(II) salt such aspalladium(II)acetate under an atmosphere of carbon monoxide will lead tothe ester IX. The hydrolysis of the ester IX may be achieved using aninorganic base such as lithium hydroxide in water and the resulting acidmay be reduced to the alcohol XI by treatment with a chloroformate suchas isopropyl chloroformate in the presence of an organic base such astriethylamine in an organic solvent such as THF, followed by addition ofa reducing agent such as sodium borohydride in water. The alcohol XI maybe then converted to the halide XII by treatment withtriphenylphosphine, imidazole and CBr₄ in an organic solvent such as CH₂Cl₂. Coupling of halide XII with the appropriate phenol XIII in anorganic solvent such as DMF using an inorganic base such as K₂ CO₃provides compounds of formula I (wherein X³ =--OCH₂ --) of the presentinvention.

Scheme IV

Scheme IV illustrates the conversion of compounds of formula I (whereinR¹¹ /R¹² =H) to formula I (wherein R¹¹ /R¹² is not H). Compounds offormula I (wherein R¹¹ /R¹² =H) may be alkylated by treatment with anorganic base such as lithium diisopropylamide in an organic solvent suchas THF, followed by quenching with an alkyl halide such as methyliodide. A second alkylation may be achieved by using the sameconditions. Conversion of compounds of formula I (wherein R¹¹ /R¹² =H)to compounds I (wherein R¹¹ =OH, R¹² =H) may be achieved by anhydrolysis process using an inorganic base such as NaOH in a solventsuch as EtOH/H₂ O to provide the carboxylate salt XIV, followed by anoxidation step using an oxidizing agent such as pyridiniumchlorochromate in an organic solvent such as CH₂ Cl₂. The reaction ofcompound I (R¹¹ =OH, R¹² =H) with a lower alkyl Grignard or lithiumreagent may yield compound I (R¹¹ =lower alkyl, R¹² =H). ##STR9##

Representative Compounds

Table I illustrates compounds representative of the present invention.

                  TABLE I                                                         ______________________________________                                         ##STR10##                     Ib                                             EX         R.sup.1                                                                             R.sup.3    R.sup.7                                                                            Het                                          ______________________________________                                        1          H     H          OH   5,3-Pye                                      2          H     H          OMe  5,3-Pye                                      3          H     H          OH   6,2-Pye                                      4          H     H          OMe  6,2-Pye                                      5          H     H          OH   4,2-Pye                                      6          H     H          OMe  4,2-Pye                                      7          CH.sub.2 O       OH   5,3-Pye                                      8          CH.sub.2 O       OMe  5,3-Pye                                      9          CH.sub.2 O       OH   6,2-Pye                                      10         CH.sub.2 O       OMe  6,2-Pye                                      11         CH.sub.2 O       OH   2,4-Pye                                      12         CH.sub.2 O       OMe  2,4-Pye                                      ______________________________________                                    

Assays for Determining Biological Activity

Compounds of Formula I can be tested using the following assays todetermine their mammalian leukotriene biosynthesis inhibiting activity.

Human 5-Lipoxygenase Inhibitor Screen

Objective of the Assay: The objective of the assay is to select agentswhich specifically inhibit the activity of human 5-lipoxygenase using a100,000×g supernatant fraction prepared from insect cells infected withrecombinant baculovirus containing the coding sequence for human5-lipoxygenase. Enzyme activity is measured spectrophotometrically fromthe optimal rate of conjugated diene formation (A₂₃₄) measured after theincubation of the enzyme with arachidonic acid in the presence of ATP,calcium ions and phosphatidylcholine.

Description of Procedure: The activity of 5-lipoxygenase is measuredusing a spectrophotometric assay and recombinant human 5-lipoxygenase asa source of enzyme. The 100,000×g fraction from S19 cells infected withthe recombinant baculovirus rvH5LO(8-1) containing the coding regionsequence for human 5-lipoxygenase is prepared as described by Denis etal. (J. Biol. Chem., 266, 5072-5079 (1991)). The enzymatic activity ismeasured, using a spectrophotometric assay from the optimal rate ofconjugated diene formation (A₂₃₄) using the procedure described byRiendeau et al. (Blochem. Pharmacol. 38, 2323-2321, (1989)) with minormodifications. The incubation mixture contains 50 mM sodium phosphate pH7.4, 0.2 mM ATP, 0.2 mM CaCl₂, 20 μM arachidonic acid (5 μL from a100-fold concentrated solution in ethanol), 12 μg/mLphosphatidylcholine, an aliquot of the 100,000×g fraction (2-10 μL) andinhibitor (0.5 mL final volume). Inhibitors are added as 500-foldconcentrated solutions in DMSO. Reactions are initiated by the additionof an aliquot of the enzyme preparation and the rate of conjugated dieneformation is followed for 2 minutes at room temperature. The reactionsare performed in semi-micro cuvettes (0.7 mL capacity, 10 mm path lengthand 4 mm internal width) and the absorbance changes are recorded with aHewlett-Packard diode array spectrophotometer (HP 8452A) connected tothe ChemStation using UV/VIS Kinetics Software (Hewlett-Packard).Enzymatic activity is calculated from the optimal rate of the reactionby a linear fit of the variation of A₂₃₄ during the first twenty secondsusing the least square method for the equation A.sub. 234 =V_(o) t+A_(o)where V_(o) is the rate, t is the time, and A_(o) is the absorbance atzero time. The results are expressed as percentages of inhibition of thereaction rate relative to controls (typically between 0.15-0.21AU/min)containing the DMSO vehicle.

Rat Peritoneal Polymorphonuclear (PMN) Leukocyte Assay

Rats under ether anesthesia are injected (i.p.) with 8 mL of asuspension of sodium caseinate (6 grams in ca. 50 mL water). After 15-24hr. the rats are sacrificed (CO₂) and the cells from the peritonealcavity are recovered by lavage with 20 mL of buffer (Eagles MEMcontaining 30 mM HEPES adjusted to pH 7.4 with NaOH). The cells arepelleted (350×g, 5 min.), resuspended in buffer with vigorous shaking,filtered through lens paper, recentrifuged and finally suspended inbuffer at a concentration of 10 cells/mL. A 500 μL aliquot of PMNsuspension and test compound are preincubated for 2 minutes at 37° C.,followed by the addition of 10 μM calcium ionophore A-23187. Thesuspension is stirred for an additional 4 minutes then bioassayed forLTB₄ content by adding an aliquot to a second 500 μL portion of the PMNat 37° C. The LTB₄ produced in the first incubation causes aggregationof the second PMN, which is measured as a change in light transmission.The size of the assay aliquot is chosen to give a submaximaltransmission change (usually -70%) for the untreated control. Thepercentage inhibition of LTB₄ formation is calculated from the ratio oftransmission change in the sample to the transmission change in thecompound-free control.

Human Polymorphonuclear (PMN) Leukocyte LTB₄ Assay

A. Preparation of Human PMN. Human blood is obtained by antecubitalvenepuncture from consenting volunteers who have not taken medicationwithin the previous 7 days. The blood is immediately added to 10% (v/v)trisodium citrate (0.13M) or 5% (v/v) sodium heparin (1000 IU/mL). PMNsare isolated from anticoagulated blood by dextran sedimentation oferythrocytes followed by centrifugation through Ficoll-Hypaque (specificgravity 1.077), as described by Boyum (Scand. J. Clin. Lab. Invest., 21(Supp 97), 77 (1968)). Contaminating erythrocytes are removed by lysisfollowing exposure to ammonium chloride (0.16M) in Tris buffer (pH7.65), and the PMNs are resuspended at 5×10⁵ cells/mL in HEPES (15mM)-buffered Hanks balanced salt solution containing Ca²⁺ (1.4 mM) andMg²⁺ (0.7 mM), pH 7.4.

B. Generation and Radioimmunoassay of LTB₄. PMNs (0.5 mL; 2.5×105 cells)are placed in plastic tubes and incubated (37° C., 2 min) with testcompounds at the desired concentration or vehicle (DMSO, finalconcentration 0.2%) as control. The synthesis of LTB₄ is initiated bythe addition of calcium ionophore A23187 (final concentration 10 +μM) orvehicle in control samples and allowed to proceed for 5 minutes at 37°C. The reactions are then terminated by the addition of cold methanol(0.25 mL) and samples of the entire PMN reaction mixture are removed forradioimmunoassay of LTB₄.

Samples (50 μL) of authentic LTB₄ of known concentration inradioimmunoassay buffer (RIA) buffer (potassium phosphate 1 mM; disodiumEDTA 0.1 mM; Thimerosal 0.025 mM; gelatin 0.1%, pH 7.3) or PMN reactionmixture diluted 1:1 with RIA buffer are added to reaction tubes.Thereafter [³ H]-LTB₄ (10 nCi in 100 μL RIA buffer) and LTB4-antiserum(100 μL of a 1:3000 dilution in RIA buffer) are added and the tubesvortexed. Reactants are allowed to equilibrate by incubation overnightat 4° C. To separate antibody-bound from free LTB₄, aliquots (50 μL) ofactivated charcoal (3% activated charcoal in RIA buffer containing 0.25%Dextran T-70) are added, the tubes vortexed, and allowed to stand atroom temperature for 10 minutes prior to centrifugation (1500×g; 10 min;4° C.). The supernatants containing antibody-bound LTB₄ are decantedinto vials and Aquasol 2 (4 mL) is added. Radioactivity is quantified byliquid scintillation spectrometry. The specificity of the antiserum andthe sensitivity of the procedure have been described by Rokach et al.,Prostaglandins Leukotrienes and Medicine, 13, 21 (1984). The amount ofLTB₄ produced in test and control samples is calculated. Inhibitorydose-response curves are constructed using a four-parameter algorithmand from these the IC₅₀ values are determined.

Human Whole Blood Assay In Vitro for LTB₄ Production

Fresh blood is collected in heparinized tubes by venipuncture from humanvolunteers. A 500 μL aliquot is incubated with one of the test compoundsat final concentrations varying from 3 nM to 3 mM at 37° C. for 15 min.Drug stock solutions are made up in DMSO and 1 μL of the stock solutionis added to each assay tube. The blood is then incubated with A23187 (in5 μL autologous plasma, 25 pM final concentration) at 37° C. for 30 min.At the end of incubation, plasma is obtained (12,000×g, 15 min) and a100 μL aliquot is added to 400 μL methanol for protein precipitation.The mixture is vortexed, centrifuged and the supernatant stored at -70°C. until assayed for LTB₄ by standard RIA.

Asthmatic Rat Assay

Rats are obtained from an inbred line of asthmatic rats. Both female(190-250 g) and male (260-400 g) rats are used.

Egg albumin (EA), grade V, crystallized and lyophilized, is obtainedfrom Sigma Chemical Co., St. Louis. Aluminum hydroxide is obtained fromthe Regis Chemical Company, Chicago. Methysergide bimaleate is suppliedby Sandoz Ltd., Basel.

The challenge and subsequent respiratory recordings are carried out in aclear plastic box with internal dimensions 10×6×4 inches. The top of thebox is removable; in use, it is held firmly in place by four clamps andan airtight seal is maintained by a soft rubber gasket. Through thecenter of each end of the chamber a DeVilbiss nebulizer (No. 40) isinserted via an airtight seal and each end of the box also has anoutlet. A Fleisch No. 0000 pneumotachograph is inserted into one end ofthe box and coupled to a Grass volumetric pressure transducer (PT5-A)which is then connected to a Buxco Electronics preamplifier (BuxcoElectronics Inc., Sharon, Conn.). The preamplifier is connected to aBeckman Type R Dynograph and to a Buxco computer consisting of waveformanalyser, Data Acquisition Logger with special software. Whileaerosolizing the antigen, the outlets are open and the pneumotachographis isolated from the chamber. The outlets are closed and thepneumotachograph and the chamber are connected during the recording ofthe respiratory patterns. For challenge, 2 mL of a 3% solution ofantigen in saline is placed into each nebulizer and the aerosol isgenerated with air from a small Potter diaphragm pump operating at 10psi and a flow of 8 liters/minute.

Rats are sensitized by injecting (subcutaneously) 1 mL of a suspensioncontaining 1 mg EA and 200 mg aluminum hydroxide in saline. They areused between days 12 and 24 post sensitization. In order to eliminatethe serotonin component of the response, rats are pretreatedintravenously 5 minutes prior to aerosol challenge with 3.0 μg/kg ofmethysergide. Rats are then exposed to an aerosol of 3% EA in saline forexactly 1 minute, then their respiratory profiles are recorded for afurther 30 minutes. The duration of continuous dyspnea is measured bythe Buxco computer.

Compounds are generally administered either orally 2-4 hours prior tochallenge or intravenously 2 minutes prior to challenge. They are eitherdissolved in saline or 1% methocel or suspended in 1% methocel. Thevolume injected is 1 mL/kg (intravenously) or 10 mL/kg (orally). Priorto oral treatment rats are starved overnight. The activity of compoundsis determined in terms of their ability to decrease the duration ofantigen-induced dyspnea in comparison with a group of vehicle-treatedcontrols. Usually, a compound is evaluated at a series of doses and anED50 is determined. This is defined as the dose (mg/kg) which wouldinhibit the duration of symptoms by 50%.

Pulmonary Mechanics in Trained Conscious Squirrel Monkeys

The test procedure involves placing trained squirrel monkeys in chairsin aerosol exposure chambers. For control purposes, pulmonary mechanicsmeasurements of respiratory parameters are recorded for a period ofabout 30 minutes to establish each monkey's normal control values forthat day. For oral administration, compounds are dissolved or suspendedin a 1% methocel solution (methylcellulose, 65HG, 400 cps) and given ina volume of 1 mL/kg body weight. For aerosol administration ofcompounds, a DeVilbiss ultrasonic nebulizer is utilized. Pretreatmentperiods vary from 5 minutes to 4 hours before the monkeys are challengedwith aerosol doses of either leukotriene D₄ (LTD₄) or Ascaris suumantigen, 1:25 dilution.

Following challenge, each minute of data is calculated by computer as apercent change from control values for each respiratory parameterincluding airway resistance (R_(L)) and dynamic compliance (C_(dyn)).The results for each test compound are subsequently obtained for aminimum period of 60 minutes post challenge which are then compared topreviously obtained historical baseline control values for that monkey.In addition, the overall values for 60 minutes post-challenge for eachmonkey (historical baseline values and test values) are averagedseparately and are used to calculate the overall percent inhibition ofLTD₄ or Ascaris antigen response by the test compound. For statisticalanalysis, paired t-test is used. (References: McFarlane, C. S. et al.,Prostaglandins, 28, 173-182 (1984) and McFarlane, C. S. et al., AgentsActions, 22, 63-68 (1987).)

Prevention of Induced Bronchoconstriction in Allergic Sheep

A. Rationale. Certain allergic sheep with known sensitivity to aspecific antigen (Ascaris suum) respond to inhalation challenge withacute and late bronchial responses. The time course of both the acuteand the late bronchial responses approximates the time course observedin asthmatics and the pharmacological modification of both responses issimilar to that found in man. The effects of antigen in these sheep arelargely observed in the large airways and are conveniently monitored aschanges in lung resistance or specific lung resistance.

B. Methods. Animal Preparation: Adult sheep with a mean weight of 35 kg(range, 18 to 50 kg) are used. All animals used meet two criteria: a)they have a natural cutaneous reaction to 1:1,000 or 1:10,000 dilutionsof Ascaris suum extract (Greer Diagnostics, Lenois, N.C.) and b) theyhave previously responded to inhalation challenge with Ascaris suum withboth an acute bronchoconstriction and a late bronchial obstruction (W.M. Abraham, et al., Am. Rev. Resp. Dis., 128, 839-44 (1983)).

Measurement of Airway Mechanics: The unsedated sheep are restrained in acart in the prone position with their heads immobilized. After topicalanesthesia of the nasal passages with 2% lidocaine solution, a ballooncatheter is advanced through one nostril into the lower esophagus. Theanimals are then intubated with a cuffed endotracheal tube through theother nostril using a flexible fiberoptic bronchoscope as a guide.Pleural pressure is estimated with the esophageal balloon catheter(filled with one mL of air), which is positioned such that inspirationproduces a negative pressure deflection with clearly discerniblecardiogenic oscillations. Lateral pressure in the trachea is measuredwith a sidehole catheter (inner dimension, 2.5 mm) advanced through andpositioned distal to the tip of the nasotracheal tube. Transpulmonarypressure, the difference between tracheal pressure and pleural pressure,is measured with a differential pressure transducer (DP45; ValidyneCorp., Northridge, Calif.). For the measurement of pulmonary resistance(R_(L)), the maximal end of the nasotrachel tube is connected to apneumotachograph (Fleisch, Dyna Sciences, Blue Bell, Pa.). The signalsof flow and transpulmonary pressure are recorded on an oscilloscope(Model DR-12; Electronics for Medicine, White Plains, N.Y.) which islinked to a PDP-11 Digital computer (Digital Equipment Corp., Maynard,Mass.) for on-line calculation of R_(L) from transpulmonary pressure,respiratory volume obtained by integration and flow. Analysis of 10-15breaths is used for the determination of R_(L). Thoracic gas volume(V_(tg)) is measured in a body plethysmograph, to obtain specificpulmonary resistance (SR_(L) =R_(L) •V_(tg)).

Aerosol Delivery Systems: Aerosols of Ascaris suum extract (1:20) aregenerated using a disposable medical nebulizer (Raindrop®, PuritanBennett), which produces an aerosol with a mass median aerodynamicdiameter of 6.2 μM (geometric standard deviation, 2.1) as determined byan electric size analyzer (Model 3030; Thermal Systems, St. Paul,Minn.). The output from the nebulizer is directed into a plastict-piece, one end of which is attached to the nasotracheal tube, theother end of which is conected to the inspiratory part of a Harvardrespirator. The aerosol is delivered at a tidal volume of 500 mL of arate of 20 per minute. Thus, each sheep receives an equivalent dose ofantigen in both placebo and drug trials.

Experimental Protocol: Prior to antigen challenge baseline measurementsof SR_(L) are obtained, infusion of the test compound is started 1 hrprior to challenge, the measurement of SR_(L) repeated and then thesheep undergoes inhalation challenge with Ascaris suum antigen.Measurements of SR_(L) are obtained immediately after antigen challengeand at 1, 2, 3, 4, 5, 6, 6.5, 7, 7.5, and 8 hrs after antigen challange.Placebo and drug tests are separated by at least 14 days. In a furtherstudy, sheep are given a bolus dose of the test compound followed by aninfusion of the test compound for 0.5-1 hr prior to Ascaris challengeand for 8 hrs after Ascaris as described above.

Statistical Analysis: A Kruskal-Wallis one way ANOVA test is used tocompare the acute immediate responses to antigen and the peak lateresponse in the controls and the drug-treated animals.

PREPARATION OF ALCOHOLS Alcohol 1:5-[4-(4-Hydroxy)tetrahydropyranyl]pyridin-3-ylmethanol ##STR11## Step 1:5-Bromo-O-tert-butyldiphenylsilylpyridin-3-ylmethanol

To a solution of 5-bromopyridin-3-ylmethanol (Chem. Pharm. Bull. 1990,38, 2446) (29 g, 154 mmoL) and tert-butylchlorodiphenylsilane (47.5 g,173 mmoL) in CH₂ Cl₂ (500 mL) at r.t., there was added imidazole (15.8g, 232 mmoL). The mixture was stirred for 1 hr. and filtered. Thefiltrate was evaporated and the residue chromatographed on silica geleluting with a 1:7 mixture of EtOAc and hexane, to afford the product asa colorless oil.

Step 2:5-[4-(4-Hydroxy)tetrahydropyranyl]-O-tert-butyldiphenylsilylpyridin-3-ylmethanol

To a solution of the silylether from Step 1 (50 g, 117 mmoL) in THF (500mL), cooled to -70° C., there was slowly added n-BuLi 1.12M in hexanes(115 mL, 129 mmoL) affording a dark brown solution. To this, there wasadded a solution of tetrahydro-4H-pyran-4-one (14.1 g, 141 mmoL) in THF(925 mL). The resulting mixture was stirred for 1 hr. at -70° C., thenquenched slowly with saturated aqueous NH₄ Cl (50 mL) and allowed towarm up to r.t. After diluting with EtOAc (500 mL) the mixture waswashed (4×) with brine, dried over Na₂ SO₄, and evaporated.Chromatography on silica gel, eluting with EtOAc, afforded the productas an oil which solidified.

Step 3: 5-[4-(4-Hydroxy)tetrahydropyranyl]pyridin-3-ylmethanol

To a solution of the silylether from Step 2 (20.35 g, 45.5 mmoL) in THF(350 mL), there was added Bu₄ NF 1M in THF (52 mL) and the mixture wasstirred at r.t. for 1 hr. The solvent was evaporated and the residuechromatographed as a short column of silica gel, eluting with a 1:4mixture of EtOH and EtOAc to afford the title product which wasobtained, after trituration with Et₂ O and filtration, as a light yellowsolid; m.p. 145°-147° C.

Alcohol 2: 6-[4-(4-Hydroxy )tetrahydropyranyl]-pyridin-2-ylmethanol##STR12## Step 1: 2-Bromo-6[4-(4-hydroxy)tetrahydropyranyl]-pyridine

A solution of 2,6-dibromopyridine (15 g) in Et₂ O (375 mL) was cooled to-78° C. To the resulting suspension was slowly added n-BuLi 2M inhexanes (47.5 mL, 0.9 eq.) and the resulting mixture was stirred for afurther 15 min. at -78° C. There was slowly added a solution oftetrahydro-4H-pyran-4-one (11.6 g) in Et₂ O (25 mL). The resulting whitesuspension was stirred at -78° C. for an additional 15 min. There wasadded saturated aqueous NH₄ Cl (100 mL) and the mixture was allowed towarm up to r.t. After dilution with EtOAc, the organic phase was washed(4×) with brine, dried, and evaporated. The residue was triturated withEt₂ O and filtered to afford the title product as a white solid; m.p.131°-133° C.

Step 2: 6-[4-(4-Hydroxy)tetrahydropyranyl]pyridin-2-ylmethanol

To a solution of the bromo derivative from Step 1 (7.7 g) in THF (50 mL)and Et₂ O (150 mL), cooled to 0° C., there was slowly added n-BuLi 2M inhexanes (30 mL) affording a red-brown suspension. An inlet tube abovethe surface of the mixture was connected to a flask in whichparaformaldehyde (25 g) was gently heated at 175° C. to generateformaldehyde. When all the paraformaldehyde had been decomposed, to thereaction mixture was added saturated aqueous NH₄ Cl (100 mL) and EtOAc(500 mL). The organic phase was washed (4×) with brine, dried, andevaporated to a residue which was chromatographed on silica gel, elutingwith EtOAc to afford the title product as a thick yellow oil.

Alcohol 3: 6-[4-(4-Methoxy)tetrahydropyranyl]-pyridine-2-ylmethanol##STR13## Step 1: 2-Bromo-6-[4-(4-methoxy)tetrahydropyranyl]-pyridine

To a suspension of KH (35% dispersion in oil, 1.25 g) in THF (75 mL),cooled to 0° C., there was added2-bromo-6-[4-(4-hydroxy)tetrahydropyranyl]-pyridine from Alcohol 2,Step 1. When gassing had subsided, the mixture was warmed to r.t. and athick suspension resulted. To this was added methyl iodide (1.71 g) andthe resulting suspension was stirred at r.t. for 30 min. The THF wasevaporated away, and the residue was partitioned between H₂ O and EtOAc.The residue from evaporation of the organic phase was triturated withhexane and filtered to afford the product as a white solid; m.p. 69°-71°C.

Step 2: 6-[4-(4-Methoxy)tetrahydropyranyl]pyridin-2-ylmethanol

Following the procedure described in Alcohol 2, Step 2, but substitutingthe bromo derivative from Step 1 for2-bromo-6-[4-(4-hydroxy)tetrahydropyranyl]-pyridine, the title productwas obtained as a white solid; m.p. 84°-86° C.

Alcohol 4: 4-[4-(4-Hydroxy)tetrahydropyranyl]-pyridin-2-ylmethanol##STR14##

Following the procedure described in Alcohol 1, Steps 1-3, butsubstituting 4-bromopyridin-2-ylmethanol (Chem. Pharm. Bull. 1990, 38,2446) for 5-bromo-pyridin-3-ylmethanol as starting material, the titleproduct was obtained as a white solid.

Alcohol 5: (1S,5R)5-[3-(3α-Hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]pyridin-3-ylmethano##STR15## Step 1: 2,4-Di-O-p-toluenesulfonyl-1,6-anhydro-β-D-glucose

To a solution of 1,6-anhydro-β-D-glucose (50 g, 308 mmoL) in drypyridine (100 mL) at 0° C. was added dropwise a solution ofp-toluenesulfonyl chloride (123 g, 647 mmoL) dissolved in CHCl₃ (350 mL)and pyridine (200 mL). The reaction mixture was stirred at r.t. for atleast 2 days. Water was added and the reaction mixture was stirred for˜1 hr, then the organic layer was decanted and the aqueous phase wasreextracted with CHCl₃. The combined organic layers were washed with H₂SO₄ (10%) until the pH remained acidic, then finally washed with asaturated NH₄ OAc solution. The resulting organic layer was dried overMgSO₄ and the solvent evaporated. The syrup obtained was flashchromatographed on silica gel eluting with hexane:EtOAc (1:1) to givethe title compound an oil.

Step 2: (1S,3S,5R) 6,8-Dioxabicyclo[3.2,1]octan-3-ol

The ditosylate derivative from Step 1 (107 g, 0.228 mmoL) was dissolvedin THF (1.6 L) at -40° C. and Super-hydride in THF (800 mL, 1M, 0.8mmoL) was slowly added. The resulting reaction mixture was stirred atr.t. overnight. The reaction was cannulated into cold H₂ O (226 mL)using external cooling, then NaOH 3N (640 mL, 1.92 mmol) and H₂ O₂ (30%)(490 mL, 4.3 mmoL) were successively added. The reaction was stirred atr.t. for 1 hr, then the supernatant (THF layer) was separated from theaqueous layer and concentrated. The resulting residue was combined withthe aqueous layer and extracted with CH₂ Cl₂ using a continuousextractor. The organic layer was dried (MgSO₄) and evaporated todryness. The oily residue was dissolved in hot Et₂ O, filtered, andevaporated to dryness affording the title compound contaminated with the2-octanol isomer. The crude product was used as such for the next step.

Step 3: (1S,5R) 6,8-Dioxabicyclo[3.2.1]octan-3-one

The crude alcohol from Step 2 (16.6 g, 89 mmoL) in CH₂ Cl₂ (200 mL) wasadded slowly to a suspension of PCC (38.4 g, 178 mmoL) and celite (22 g)in CH₂ Cl₂ (400 mL) and stirred for 1 hr. The reaction mixture wasdiluted with Et₂ O (600 mL) and filtered over celite. The filtrate wasevaporated and the residue distilled with a Kugelrohr apparatus (100°C., 1.8 mm/Hg) affording the title product as an oil.

Step 4: (1S,5R)5-[3-(3α-Hydroxy-6,8-dioxabicyclo-[3.2.1]octanyl)]pyridin-3-ylmethanol

Following the procedure described in Alcohol 1, Steps 2-3, butsubstituting (1S,5R) 6,8-dioxabicyclo[3.2.1]octan-4-one from Step 3, fortetrahydro-4H-pyran-4-one, the title product was obtained as a whitesolid.

Alcohol 6: (1S,5R)6-[3-(3α-Hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]pyridin-2-ylmethanol##STR16## Step 1: 6-Bromo-O-tert-butyldiphenylsilylpyridin-2-ylmethanol

Following the procedure described in Alcohol 1, Step 1, but substituting6-bromopyridin-2-ylmethanol (Chem. Pharm. Bull. 1990, 38, 2446) for5-bromopyridin-3-ylmethanol, the title product was obtained as acolorless oil.

Step 2: (1S,5R)6-[3-(3α-Hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]pyridin-2-ylmethanol

Following the procedure described in Alcohol 1, Steps 2-3, butsubstituting 6-bromo-O-tert-butyldiphenylsilypyridin-2-ylmethanol fromStep 1, for 5-bromo-O-tert-butyldiphenylsilylpyridin-3-ylmethanol andsubstituting (1S,5R) 6,8-dioxabicyclo[3.2.1]-octan-4-one from Alcohol 5,Step 3, for tetrahydro-4H-pyran-4-one, the title product was obtained asa white solid.

The invention is further defined by reference to the following examples,which are intended to be illustrative and not limiting. All temperaturesare in degrees Celsius.

EXAMPLE 13-Hydroxymethyl-4-phenyl-7-[5-[4-(4-hydroxy)-tetrahydropyranyl]pyridin-3-ylmethoxy]-2-naphthoicacid, sodium salt Step 1: Benzaldehyde phenyl dithioacetal

To a solution of benzaldehyde (13.3 g) and thiophenol (26 mL) inisopropyl acetate (200 mL), cooled in an ice-water bath, there wasslowly added BF₃ •Et₂ O (15 mL). The resulting mixture was stirred inthe cold for an additional hour. There was slowly added 10% aqueous K₂CO₃ (100 mL). The phases were separated, the aqueous phase was extractedwith EtOAc. The combined organic fractions were then washed with H₂ O(3×), dried, and evaporated to an oil which was used as such.

Step 2:3-[Phenyl-bis(phenylthio)methyl]-2-(3-benzyloxy-α-hydroxybenzyl)butyrolactone

To a solution of the dithioacetal from Step 1 (5.7 g) in THF (100 mL),at -70° C. there was slowly added 2.1M n-BuLi in hexanes (9.0 mL). Theresulting dark solution was stirred for a further 20 min. at -70° C.,then there was added dropwise 2-(5H)furanone (Omega Inc. 1.6 mL), and 30min. later, a solution of 3-benzyloxybenzaldehyde (Aldrich, 2.0 g) inTHF 20 mL). The mixture was stirred a further hour at -70° C., thenthere was slowly added glacial HOAc (1.5 mL). The mixture was allowed towarm up to r.t., diluted with Et₂ O (500 mL), washed with brine (4×),dried, and evaporated. The residue was used as such in the next step.

Step 3: 3-Hydroxymethyl-4-phenyl-7-hydroxy-2-naphthoic acid, lactoneform

The crude product from Step 2 was dissolved in thioanisole (20 mL),there was added TFA (30 mL), and the mixture was stirred at r.t. for 18hr. After cooling, the TFA was evaporated, the residue was diluted withEt₂ O (100 mL) and after 20 min., the insoluble solid was filtered toafford the desired title compound.

Step 4:3-Hydroxymethyl-4-phenyl-7-[5-[4-(4-hydroxy)-tetrahydropyranyl]pyridin-3-ylmethoxy]-2-naphthoicacid, lactone form

To a mixture of lactone (238 mg) from Step 3,5-[4-(4-hydroxy)tetrahydropyranyl]pyridin-3-ylmethanol (Alcohol 1, 180mg) and triphenylphosphine (270 mg) in THF (8 mL), there was addeddi-tert-butyl azodicarboxylate (179 mg) and the mixture was stirred atr.t. for 2 hr. After evaporation of the THF, the residue waschromatographed on a column of silica gel, eluting with a 1:9 mixture ofEtOH and EtOAc. The product obtained was triturated with a 1:1 mixtureof Et₂ O and hexane, affording on filtration the title compound as acream-colored solid; m.p. 172°-174° C.

3-Hydroxymethyl-4-phenyl-7-[5-[4-(4-hydroxy)-tetrahydropyranyl]pyridin-3-ylmethoxy]-2-naphthoicacid, sodium salt

To a suspension of the lactone from Step 4 in EtOH is added 1N NaOH (1eq.) and refluxed 18 hr. The resulting solution is evaporated to affordthe title compound.

EXAMPLES 2-12

Following the procedure described in Example 1, Steps 4-5, butsubstituting the appropriate alcohol for5-[4-(4-hydroxy)tetrahydropyranyl]pyridin-3-ylmethanol the compounds ofExamples 2-12 are obtained.

What is claimed is:
 1. A compound of the formula: ##STR17## wherein: R¹, R⁵, and R¹¹ is each independently H, OH, lower alkyl, or lower alkoxy;R² is H, lower alkyl, or together with R¹ forms a double bonded oxygen (═O); R³ is H, lower alkyl, hydroxy lower alkyl, lower alkoxy lower alkyl, or is joined to R¹ to form a carbon bridge of 2 or 3 carbon atoms or a mono-oxa carbon bridge of 1 or 2 carbon atoms, said bridge optionally containing a double bond; R⁴, R¹², and R¹⁴ is each independently H or lower alkyl; R⁶ H or lower alkyl, or two R⁶ groups on the same or adjacent carbons can form a saturated ring of 3 to 8 members; R⁷ is H, OH, lower alkyl, lower alkoxy, lower alkylthio, or lower alkylcarbonyloxy; R⁸ is H, halogen, lower alkyl, lower alkoxy, CF₃, CN, or COR¹⁴ ; R⁹ and R¹⁰ is each independently H, lower alkyl, lower alkoxy, hydroxy lower alkyl, lower alkoxy lower alkyl, lower alkylthio lower alkyl, lower alkythio lower alkylcarbonyl, (R⁸)₂ -phenylthio lower alkyl, halogen, CN, NO₂, CF₃, N₃, N(R¹³)₂, NR¹³ COR¹⁴, NR¹³ CON(R¹³)₂, SR¹⁵, S(O)R¹⁵, S(O)₂ R¹⁵, S(O)₂ N(R¹³)₂, COR¹⁴, CON(R¹³)₂, CO₂ R¹⁴, C(R¹⁴)₂ OC(R¹⁴)₂ --CO₂ R¹⁴, or C(R¹⁴)₂ CN; R¹⁰ is attached to either ring of the naphthalene ring system; R¹³ H or lower alkyl, or two R¹³ groups attached to the same nitrogen may form a saturated ring of 5 or 6 members, optionally containing a second heteroatom chosen from O, S, or NR⁴ ; R¹⁵ is lower alkyl, phenyl-(R⁸)₀₋₂, or CF₃ ; X¹ is O, S, S(O), S(O)₂, or C(R⁶)₂ ; X² is O, S, C(R⁶)₂, or bond; X³ is C(R⁶)₂ S, SC(R⁶)₂, C(R⁶)₂ O, OC(R⁶)₂, CR⁶ ═CR⁶, C(R⁶)₂ C(R⁶)₂, O, or S; Ar is phenyl(R⁹)₂ or naphthyl(R⁹)₂ ; Het is arylene-(R⁸)₀₋₂, wherein arylene is a 5-membered aromatic ring wherein one carbon atom is replaced by O or S and 0-2 carbon atoms are replaced by N; a 5-membered aromatic ring where 1-3 carbon atoms are replaced by N; a 6-membered aromatic ring wherein 1-3 carbon atoms are replaced by N; 2- or 4-pyranone; or 2- or 4-pyridinone; m is 0 or 1;or a pharmaceutically acceptable salt thereof.
 2. A compound of claim 1 of the formula: ##STR18## or a pharmaceutically acceptable salt thereof.
 3. A compound of the formula: ##STR19## wherein: R¹ is H, Me, OMe;R³ is H or Me; R¹ and R³ together are --CH═CH--, --CH₂ CH₂ --, --OCH₂ --, or --CH₂ O--; R⁷ is OH or OMe; and Het is 5,3-Pye; 6,2-Pye; 4,2-Pye, or 2,4-Pye;or a pharmaceutically acceptable salt thereof.
 4. A compound of claim 3 of the formula ##STR20## wherein the substituents are as follows:

    ______________________________________                                         R.sup.1 R.sup.3       R.sup.7                                                                               Het                                               ______________________________________                                         H       H             OH     5,3-Pye                                           H       H             OMe    5,3-Pye                                           H       H             OH     6,2-Pye                                           H       H             OMe    6,2-Pye                                           H       H             OH     4,2-Pye                                           H       H             OMe    4,2-Pye                                           --CH.sub.2 O--        OH     5,3-Pye                                           --CH.sub.2 O--        OMe    5,3-Pye                                           --CH.sub.2 O--        OH     6,2-Pye                                           --CH.sub.2 O--        OMe    6,2-Pye                                           --CH.sub.2 O--        OH     2,4-Pye                                           --CH.sub.2 O--        OMe    2,4-Pye                                           ______________________________________                                    


5. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier.
 6. A pharmaceutical composition of claim 5 additionally comprising an effective amount of second active ingredient selected from the group consisting of non-steroidal anti-inflammatory drugs; peripheral analgesic agents; cyclooxygenase inhibitors; leukotriene antagonists; leukotriene biosynthesis inhibitors; H₁ - or H₂ -receptor antagonists; antihistaminic agents; prostaglandin antagonists; thromboxane antagonists; thromboxane synthetase inhibitors; and ACE antagonists.
 7. A pharmaceutical composition of claim 6, wherein the second active ingredient is a non-steroidal anti-inflammatory drug.
 8. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1, an effective amount of a second active ingredient which is a non-steroidal anti-inflammatory drug, and a pharmaceutically acceptable carrier, wherein the weight ratio of said compound of claim 1 to said second active ingredient ranges from about 1000:1 to 1:1000. 